Abstract:Abstract: It has been confirmed that bone marrow mesenchymal stem cells have the potential to differentiate into hepatocytes, however, the related regulation mechanism has not been elucidated. To explore the specific mechanism, OCT4 gene promoter methylation variation was detected in differentiation induction of rat bone marrow-derived liver stem cells (RBMLSCs) into hepatocytes in vitro. RBMLSCs were induced by 25 μg/L recombinant human hepatocyte growth factor and 0.1 nmol/L dexamethasone. DNA and RNA were extracted from cells at different time points (day 0, 7 and 14) after induction. Fluorescence quantitative PCR (FQ-PCR) was performed to detect OCT4 and albumin (Alb) mRNA expression. Pyrosequencing was performed to detect methylation variation of 4 CpG sites in OCT4 promoter. During cell differentiation, Alb mRNA expression was increased significantly (P<0.05), but OCT4 mRNA was decreased significantly (P<0.005) at day 7 and day 14. Methylation frequency of the first 3 CpG sites in OCT4 promoter upstream was increased significantly (P<0.001), but the 4th CpG site had no obvious methylation change. The results showed that the hypermethylation of OCT4 promoter may lead to the decline of its mRNA expression, disappearance of RBMLSCs pluripotency, but that the 4th CpG site might not be involved in the regulating process.
引用本文:
潘润华, 阿斯楞, 张 茂, 利 民, 张梓朗, 廖彩仙. 焦磷酸测序法检测OCT4启动子在大鼠骨髓源性肝干细胞分化中的甲基化变化[J]. 生命科学研究, 2016, 20(2): 113-118.
PAN Run-Hua, A Si-Leng, ZHANG Mao, LI Min, ZHANG Zi-Lang, LIAO Cai-Xian. Pyrosequencing Detection of OCT4 Promoter Methylation in Differentiation of Rat Bone Marrow-derived Liver Mesenchymal Stem Cells. Life Science Research, 2016, 20(2): 113-118.